生物技术通报2017，Vol.33Issue(3)：186-192,7.DOI:10.13560/j.cnki.biotech.bull.1985.2017.03.027
LAMP技术快速诊断布鲁氏菌病的研究
Rapid Diagnosis of Brucella by Loop-mediated Isothermal Amplification
摘要
Abstract
This study aims to apply the loop-mediated isothermal amplification(LAMP)in detecting Brucella. A set of six specific primers specific to regions of 16S rDNA gene were designed,and the turbidity technique was employed to optimize the reaction conditions,by which the specificity and sensitivity of the method were evaluated. Results are :(1)the optimized temperature was 62℃ at constant 60 min, and the concentration of inner primer was 1.50 μmol/μL,0.20 μmol/μL outer primers,and 1.0 μmol/μL loop primers ;(2)furthermore, 3 strains of Brucella occurred LAMP amplification reaction and achieved the positive turbidity value,but there was no amplification with other control group including Yersinia enterocolitica ATCC9510,Escherichia coil ATCC25922,Salmonella typhimurium ATCC10708,and Staphylococcus aureus ACTT33591 ;(3)the minimum thresholds for turbidity technique and real-time fluorescence were 4.36 fg/μL and 436 fg/μL respectively,their sensitivities were both higher than the value by conventional PCR,4.36 pg/μL. Obviously,it is fast,efficient,and convenient while LAMP is applied in the clinic diagnosis of Brucella,therefore,it is practically applicable for the pathogenic diagnosis of Brucella in grassroots communities.关键词
布鲁氏菌/16SrDNA/环介导等温扩增技术(LAMP)Key words
Brucella/16S rDNA/loop-mediated isothermal amplification(LAMP)引用本文复制引用
谢文萍,肇慧君,张琳,姜红旭,吴斌,孙浩..LAMP技术快速诊断布鲁氏菌病的研究[J].生物技术通报,2017,33(3):186-192,7.基金项目
国家质量监督检验检疫总局科研项目(2014IK248) （2014IK248）
